Biotin Conjugation Kit

Equilibrate all materials and prepared reagents to room temperature prior to use. Note: If the protein contains free amino groups (Tris, amino acids or other interferents), please centrifuge it 2-3 times (8,000 x g, 10 min) using the Filtration Tube to ensure that it is removed. For 1 mg antibody 1. Add 200 μl of Reaction Buffer into each 1 ml of antibody (e.g. antibody concentration 1 mg/ml) to be labeled and mix gently. (Reduce the amount of Reaction Buffer at the same scale) 2. Add 30 μl of 10 mM NHS Biotin into the protein solution, gently blow and mix fully. Replace cap on the vial and leave standing for 30 minutes in the dark at 37°C hot air circulating oven. 3. After incubating, add 5 μl of Quencher Reagent for each 30 μl of 10 mM NHS Biotin used and mix gently (Reduce the amount of Quencher Reagent at the same scale) . The conjugates can be used after 10 minutes. 4. Add an appropriate amount of 1xPBS into the above reaction solution, cover the cap after matching with the collection tube, and centrifuge for 10-30 min at the speed of 12,000 x g. Discard the liquid in the collection tube, replenish 1xPBS in the filter device, and repeat the centrifugal ultrafiltration operation for 2-3 times. 5. Invert the filter device in another collection tube and centrifuge at 1000 x g for 2 mins. Collect the solution in the collection tube.